ThegoalsofourR24projectincludeproducingnewrevertibleallelestoidentifygenes capableofpromotinghumanhealthandgeneratingnewmethodologiesforsite-directedsomatic mutagenesis,enhancinghomologousrecombinationandsingle-strandannealingmediated integrationofDNA.Itisanticipatedthatthesetechnologieswillgreatlyalleviatethenumberof animalsrequiredforgeneticstudiesinzebrafishandcanbeextendedtootheranimalspecies. Thesuccessofthistechnologydevelopmenthingesontheabilitytoscreenembryosandlarvae forfluorescentreportsofgeneexpressionandtransgenesis.InthisR24revision,weare proposingtoincreaseouropticalsectioningabilitiesandupdateexistingfluorescence microscopestomeettheincreaseddemandsonourimagingequipmentthatisacriticalpartof ourexperimentalstrategiesandthoseofothergroupsatIowaStateUniversity(ISU).These goalswillbeaccomplishedinthefollowingaims:Aim1.Improveopticalsectioningcapabilities oflivingzebrafishembryosatIowaState.WeareproposingtopurchaseaZeissAxioImagerZ2 equippedwithanApotome2tomeettheincreasedopticalsectioningdemandsatISU.Aim2. UpdateexistingmicroscopesusedinassociationwithourzebrafishfacilitieswithLEDlight sources.Together,theupdatesproposedabovewouldgreatlyincreaseourfluorescence screeningandimagingneedsofourcurrentlyfundedR24project,ourcolleaguesatISU,and futureinitiatives.